The research goals in the Membrane Structure and Function Section are directed towards an understanding of mechanisms of membrane fusion mediated by viral spike glycoproteins. We are specifically studying the mode of action of the G protein of Vesicular Stomatitis Virus (VSV), the HN and F proteins of Sendai Virus, and the HA protein of influenza virus. Specific topics include: i) development of fluorescent methods to study kinetics and extent of adhesion and fusion using intact and reconstituted virions, and liposomes and cells as targets; ii) development of methods to analyze reconstitution of viral spike glycoproteins; iii) functional reconstitution of viral spike glycoproteins into lipid vesicles iv) studies of mechanism based on an allosteric model: the role of ligand binding, conformational changes and cooperativity of viral spike glycoproteins in mediating membrane fusion v) studies of the effects of modifications of viral spike glycoproteins by pH temperature, enzymes, and chemicals on their fusogenic activities vi) studies of the relationship between virus- induced membrane destabilization (permeability changes) and fusion vii) Studies of viral entry into the cell by endocytosis using fluorescent techniques. viii) Application of image processing using video-enhanced fluorescent microscopy controlled by a computer to analyze viral entry pathways. ix) Examination of the disposition of the fusion protein after the fusion event; x) Identification of possible fusion intermediates; xi) Development of methods to study fusion activity of mutants of viral proteins using cloned viral membrane protein sequences expressed in transfected cells; xii) Structural studies of viral proteins; xiii) Development of methods for using reconstituted viral envelopes as vehicles for specific delivery of materials into cells.